The structure and activities of the archaeal transcription termination factor Eta detail vulnerabilities of the transcription elongation complex.

Transcription must be properly regulated to ensure dynamic gene expression underlying growth, development, and response to environmental cues. Regulation is imposed throughout the transcription cycle, and while many efforts have detailed the regulation of transcription initiation and early elongation, the termination phase of transcription also plays critical roles in regulating gene expression. Transcription termination can be driven by only a few proteins in each domain of life. Detailing the mechanism(s) employed provides insight into the vulnerabilities of transcription elongation complexes (TECs) that permit regulated termination to control expression of many genes and operons. Here, we describe the biochemical activities and crystal structure of the superfamily 2 helicase Eta, one of two known factors capable of disrupting archaeal transcription elongation complexes. Eta retains a twin-translocase core domain common to all superfamily 2 helicases and a well-conserved C terminus wherein individual amino acid substitutions can critically abrogate termination activities. Eta variants that perturb ATPase, helicase, single-stranded DNA and double-stranded DNA translocase and termination activities identify key regions of the C terminus of Eta that, when combined with modeling Eta-TEC interactions, provide a structural model of Eta-mediated termination guided in part by structures of Mfd and the bacterial TEC. The susceptibility of TECs to disruption by termination factors that target the upstream surface of RNA polymerase and potentially drive termination through forward translocation and allosteric mechanisms that favor opening of the clamp to release the encapsulated nucleic acids emerges as a common feature of transcription termination mechanisms.

Governance and the mangrove commons: Advancing the cross-scale, nested framework for the global conservation and wise use of mangroves.

Mangroves provide critical ecosystems services, contributing an estimated 42 billion US dollars to global fisheries, storing 25.5 million tons of carbon per year, and providing flood protection to over 15 million people annually. Yet, they are increasingly threatened by factors ranging from local resource exploitation to global climate change, with an estimated 35% of mangrove forests lost in the past two decades. These threats are difficult to manage due to the intrinsic characteristics of mangrove systems and their provisioning services, and their transboundary and pan-global nature. Due to their unique intertidal ecological niche, mangroves are often treated as a "common pool resource" within national legal frameworks, making them particularly susceptible to exploitation. Moreover, they form ecological connections through numerous biotic and abiotic processes that cross political boundaries. Because of these qualities a cross-scale nested framework of international, regional, and local coordination is necessary to successfully sustain mangrove ecosystems and their valuable services. Although coordination across the geopolitical spectrum is often cited as a need for effective management of common resources such as mangroves, there has been no formal analysis of mangrove multiscale governance. In this paper we address this gap by providing a comprehensive analysis of interactions between and within international, regional, and local mangrove management regimes and examine the challenges and opportunities such multiscale governance frameworks present. We highlight Costa Rica as a case study to demonstrate the universal relevance and potential of multi-scale governance and explore its downscale potential. Using Elinor Ostrom's principles for self-governance of the commons as our touchstone, we identify where improvements to the status quo could be implemented to increase its effectiveness of the current frameworks to meet the ongoing challenge of managing mangrove-derived resources and services in the face of a changing climate and human needs.

Paget's Disease of the Bone: Patterns of Referral to Secondary Care Following Diagnosis on X-rays.

Paget's disease of bone (PDB) is the second commonest metabolic bone disorder in the UK after osteoporosis and is both underdiagnosed and undertreated. PDB is often found incidentally on plain X-rays. There is effective treatment so identification of affected individuals is important. The aim was to conduct an audit to determine what proportion of individuals with X-ray evidence of PDB were referred to secondary care. A retrospective audit of X-rays reports in men and women over 55 years of age was undertaken over 18 months searching for the key word "Paget's." The images of possible cases were reviewed and the presence of PDB confirmed. The proportion already known to secondary care was determined and those that had had isotope bone scans and treatment. Data recorded included site of lesion, age, gender, level of total alkaline phosphatase (ALP) and complications. A total of 68,873 X-rays were screened and 43 cases found. Pelvic images had the highest proportion of positive findings at 0.2% and 65% of the cases. 74% had not been referred to secondary care. The mean age was 86.7 years (range 65-95) and the ALP was elevated in 65% with a mean of 189u/L (range 47-804u/L). In 33 individuals, PDB had been recorded in the reports of previous X-rays. The rate of referral for specialist care remains low. The prevalence of the condition appears to be falling.

TFS and Spt4/5 accelerate transcription through archaeal histone-based chromatin.

RNA polymerase must surmount translocation barriers for continued transcription. In Eukarya and most Archaea, DNA-bound histone proteins represent the most common and troublesome barrier to transcription elongation. Eukaryotes encode a plethora of chromatin-remodeling complexes, histone-modification enzymes and transcription elongation factors to aid transcription through nucleosomes, while archaea seemingly lack machinery to remodel/modify histone-based chromatin and thus must rely on elongation factors to accelerate transcription through chromatin-barriers. TFS (TFIIS in Eukarya) and the Spt4-Spt5 complex are universally encoded in archaeal genomes, and here we demonstrate that both elongation factors, via different mechanisms, can accelerate transcription through archaeal histone-based chromatin. Histone proteins in Thermococcus kodakarensis are sufficiently abundant to completely wrap all genomic DNA, resulting in a consistent protein barrier to transcription elongation. TFS-enhanced cleavage of RNAs in backtracked transcription complexes reactivates stalled RNAPs and dramatically accelerates transcription through histone-barriers, while Spt4-Spt5 changes to clamp-domain dynamics play a lesser-role in stabilizing transcription. Repeated attempts to delete TFS, Spt4 and Spt5 from the T. kodakarensis genome were not successful, and the essentiality of both conserved transcription elongation factors suggests that both conserved elongation factors play important roles in transcription regulation in vivo, including mechanisms to accelerate transcription through downstream protein barriers.

Quantitative Computed Tomography (QCT) of the Distal Forearm in Men Using a Spiral Whole-Body CT Scanner - Description of a Method and Reliability Assessment of the QCT Pro Software.

The Mr F study investigates the pathogenesis of low trauma distal forearm fractures in men and includes volumetric bone mineral density (vBMD) measurements at the ultradistal forearm as there are no current data. A standard 64 slice CT scanner was used to determine if it was possible to adapt the existing Mindways quantitative computed tomography Pro software for measuring vBMD values at the hip and spine sites. For calculation of intra- and interobserver reliability 40 forearm scans out of the 300 available were chosen randomly. The images were analyzed using the Slice Pick module and Bone Investigational Toolkit. The 4% length of the radius was chosen by measuring the length of the radius from the scaphoid fossa distally to the radial head. The acquired image then underwent extraction, isolation, rotation, and selection of region of interest in order to generate a report on vBMD. A cross-sectional image was created to allow the generation of data on the cortical and trabecular components separately. Repeat analyses were undertaken by 3 independent observers who were blinded as to whether the image was from a participant with or without fracture. The images were presented in random order at each time point. The following parameters were recorded: cortical cross sectional area, total vBMD, trabecular vBMD, and cortical vBMD (CvBMD). Data were analyzed by calculating intraclass correlation coefficients for intra- and interobserver reliability. The lowest values occurred at the CvBMD with intraobserver reliability of 0.92 (95% confidence interval [CI] of 0.86-0.96) and interobserver reliability of 0.92 (95% CI 0.89-0.96). All other parameters had reliability values between 0.97 and 0.99 with tighter 95% CI than for CvBMD. The method of adapting the Mindways Pro software using a standard CT to produce vBMD and structural data at the ultradistal radius is reliable.

Factor-dependent archaeal transcription termination.

RNA polymerase activity is regulated by nascent RNA sequences, DNA template sequences, and conserved transcription factors. Transcription factors promoting initiation and elongation have been characterized in each domain, but transcription termination factors have been identified only in bacteria and eukarya. Here we describe euryarchaeal termination activity (Eta), the first archaeal termination factor capable of disrupting the transcription elongation complex (TEC), detail the rate of and requirements for Eta-mediated transcription termination, and describe a role for Eta in transcription termination in vivo. Eta-mediated transcription termination is energy-dependent, requires upstream DNA sequences, and disrupts TECs to release the nascent RNA to solution. Deletion of TK0566 (encoding Eta) is possible, but results in slow growth and renders cells sensitive to DNA damaging agents. Our results suggest that the mechanisms used by termination factors in archaea, eukarya, and bacteria to disrupt the TEC may be conserved, and that Eta stimulates release of stalled or arrested TECs.

Development of a Synthetic Training Model for Canine Thoracocentesis.

Thoracocentesis, a procedure in which air or fluid is removed from the pleural space, is used to relieve respiratory distress, and as a diagnostic procedure in human and veterinary medicine. Veterinary students commonly learn and practice the procedure on canine cadavers which are in limited supply and are not amenable to long-term storage and use. Practicing thoracocentesis on a cadaveric model also provides limited feedback indicative of success and/or procedural complications. One commercial model for practicing canine thoracocentesis is available, but it costs over US$2000 and is excessively bulky. In order to improve the learning process for veterinary students, we have developed a reusable synthetic canine thorax model that accurately replicates the thoracocentesis procedure, provides immediate feedback to the students and reduces the need for canine cadavers. The low cost of our product provides an efficient alternative to cadavers for instruction in veterinary schools or hospitals.

Building a genome engineering toolbox in nonmodel prokaryotic microbes.

The realization of a sustainable bioeconomy requires our ability to understand and engineer complex design principles for the development of platform organisms capable of efficient conversion of cheap and sustainable feedstocks (e.g., sunlight, CO2 , and nonfood biomass) into biofuels and bioproducts at sufficient titers and costs. For model microbes, such as Escherichia coli, advances in DNA reading and writing technologies are driving the adoption of new paradigms for engineering biological systems. Unfortunately, microbes with properties of interest for the utilization of cheap and renewable feedstocks, such as photosynthesis, autotrophic growth, and cellulose degradation, have very few, if any, genetic tools for metabolic engineering. Therefore, it is important to develop "design rules" for building a genetic toolbox for novel microbes. Here, we present an overview of our current understanding of these rules for the genetic manipulation of prokaryotic microbes and the available genetic tools to expand our ability to genetically engineer nonmodel systems.

Differentiating between cold agglutinins and rouleaux: a case series of seven patients.

We present a case series of seven patients with suspected cold agglutinin antibodies, discovered after initiation of bypass. Laboratory analysis of blood samples intraoperatively determined the cause of the aggregation to be rouleaux formation in three of the patients and cold agglutinins in the other four.

Adult Paget's disease of bone: a review.

Adult PD of bone is the second commonest metabolic bone condition after osteoporosis. The condition is characterized by increased bone cell activity, with bone-resorbing osteoclasts often larger and containing more nuclei than normal, and osteoblasts producing increased amounts of disorganized bone. This leads to expanded bone of poor quality possessing both sclerotic and lytic areas. PD of bone has a strong genetic element, with a family history being noted in 10-20% of cases. A number of genetic defects have been found to be associated with the condition. The most common disease-associated variants identified affect the SQSTM1 gene, providing insights into disease aetiology, with the clinical value of knowledge of SQSTM1 mutation status currently under active investigation. The diagnosis may be suggested by an isolated raised total ALP without other identifiable causes. This can be confirmed on plain X-rays and the extent determined by isotope bone scan. The mainstays of treatment are the bisphosphonates, especially i.v. zoledronate, which results in long-term suppression of bone turnover. ALP is the usual means of monitoring the condition, although more specific bone turnover markers can be helpful, especially in coincident liver disease. Patients should be followed up to monitor for biochemical relapse or development of complications, which may require medical or surgical intervention.

Transcription Regulation in Archaea.

The known diversity of metabolic strategies and physiological adaptations of archaeal species to extreme environments is extraordinary. Accurate and responsive mechanisms to ensure that gene expression patterns match the needs of the cell necessitate regulatory strategies that control the activities and output of the archaeal transcription apparatus. Archaea are reliant on a single RNA polymerase for all transcription, and many of the known regulatory mechanisms employed for archaeal transcription mimic strategies also employed for eukaryotic and bacterial species. Novel mechanisms of transcription regulation have become apparent by increasingly sophisticated in vivo and in vitro investigations of archaeal species. This review emphasizes recent progress in understanding archaeal transcription regulatory mechanisms and highlights insights gained from studies of the influence of archaeal chromatin on transcription.

Analyses of in vivo interactions between transcription factors and the archaeal RNA polymerase.

Transcription factors regulate the activities of RNA polymerase (RNAP) at each stage of the transcription cycle. Many basal transcription factors with common ancestry are employed in eukaryotic and archaeal systems that directly bind to RNAP and influence intramolecular movements of RNAP and modulate DNA or RNA interactions. We describe and employ a flexible methodology to directly probe and quantify the binding of transcription factors to RNAP in vivo. We demonstrate that binding of the conserved and essential archaeal transcription factor TFE to the archaeal RNAP is directed, in part, by interactions with the RpoE subunit of RNAP. As the surfaces involved are conserved in many eukaryotic and archaeal systems, the identified TFE-RNAP interactions are likely conserved in archaeal-eukaryal systems and represent an important point of contact that can influence the efficiency of transcription initiation.

Cold Agglutinin Autoantibodies in a Patient without a Visible Coronary Sinus Ostium: Strategies for Myocardial Protection without Using Retrograde Cardioplegia.

The presence of cold agglutinins (CA) during cardiac surgery with cardiopulmonary bypass usually creates the need for an altered surgical plan. In this case, the CA were discovered after the initiation of bypass, limiting the time, and cardioplegia solutions that could be used in the new approach. The inability to cannulate the coronary sinus with a retrograde cardioplegia catheter excluded the standard approach to myocardial preservation with CA of using continuous warm blood. For this case, we used intermittent cold crystalloid delivered via the antegrade needle for the first half of the procedure and through the saphenous vein graft anastomosis during the aortic valve portion of the cross-clamp period.

Prostate cancer and osteoporosis.

Adenocarcinoma of the prostate is one of the commonest cancers in the world. Due to a combination of earlier detection and better treatments, survival has increased dramatically. Prostate cancer itself is associated with lower bone density and increased fractures. This is compounded by the use of androgen deprivation therapy, which causes dramatic falls in circulating testosterone and estrogen, resulting in rapid falls in bone density, decreased muscle mass, and increased fracture rates. Bisphosphonates have been demonstrated to prevent and reverse this bone loss, but there are no anti-fracture data. Denosumab, a monoclonal antibody to RANKL, has recently been shown to increase bone density and reduce fracture rates. Prostate cancer also commonly metastasizes to bone where it can cause complications such as fracture and pain. Both zoledronic acid and denosumab have been demonstrated to reduce skeletal related events. Comparative studies would suggest that densosumab may have an advantage over zoledronic acid.

Evaluation of the effects of corn silage maturity and kernel processing on steer growth performance and carcass traits.

Two experiments were conducted to investigate the effects of feeding kernel processed corn silage to growing calves at 65% inclusion (dry matter [DM] basis; Exp. 1] and finishing beef steers at 20% inclusion (DM basis; Exp. 2). In Exp. 1, steers (n = 184; initial shrunk body weight [BW] = 388 ± 22.3 kg) were used to evaluate the influence that kernel processing of corn silage has on production responses when fed at 65% diet inclusion (DM basis) during a 46-d growing period. Steers were allotted to 1 of 24 pens (12 replicate pens/treatment). Treatments were based upon corn silage that was either kernel processed or not. In Exp. 2, steers (n = 192; initial shrunk BW = 446 ± 28.3 kg) were used in a 112-d finishing experiment. Treatments were grouped in a 2 × 2 factorial arrangement (24 pens total; 8 steers/pen) to evaluate corn silage harvest maturity (1/2 to 2/3 milk line or black layer) and kernel processing (processed or not) at time of corn silage harvest on finishing steer growth performance and carcass traits when corn silage is fed at a dietary DM inclusion of 20%. Both experiments were analyzed as a randomized completed block design with pen as experimental unit. In Exp. 1, final BW tended (P = 0.07) to be increased by 3 kg in kernel processed corn silage. Daily weight gain and DM intake were increased (P ≤ 0.04) by 6% and 2%, respectively, in steers fed kernel processed corn silage compared to controls; however, gain efficiency was not appreciably influenced by treatment (P = 0.15). In Exp. 2, there were no harvest maturity × kernel processing interactions (P ≥ 0.26) for any growth performance measures or any parameters related to efficiency of dietary NE utilization. No harvest maturity × kernel processing interactions (P ≥ 0.08) were observed for any carcass traits except for the distribution of USDA Prime carcasses (P = 0.04). Steers fed 2/3 milk line and unprocessed corn silage had a lower (P = 0.05) proportion of carcasses grade USDA Prime (0.0%) compared to all other treatments (12.0%). Harvest time (P ≥ 0.07) and kernel processing (P ≥ 0.07) of corn silage had no appreciable influence on any other carcass trait measures. These data indicate that kernel processed corn silage fed to growing calves at 65% diet inclusion (DM basis) enhances intake and daily gain, while kernel processed corn silage fed to finishing steers at 20% diet inclusion (DM basis) does not appreciably influence daily gain, efficiency of gain, or carcass parameters.

Kernel processing of corn silage has yielded inconsistent results on diet digestibility and growth performance in beef cattle. These are likely a function of a variety of factors such as differing dry matter concentration of corn silage at harvest, diet inclusion levels, and length of cut. Two experiments were conducted to determine the effect that kernel processing of corn silage has on production responses in growing (65% dietary dry matter inclusion) and finishing beef steers (20% dietary dry matter inclusion). Data from the growing steer experiment when corn silage was included in the diet at 65% (dry matter basis) indicate that kernel processing of corn silage enhances dry matter intake and daily weight gain of beef steers with no appreciable influence on DM conversion efficiency. Data from the finishing steer experiment indicate that harvest maturity and kernel processing of corn silage have minimal effects on animal growth performance and carcass traits in finishing steers when corn silage is fed at 20% inclusion (dry matter basis). Variable responses could be related to differences in inclusion level, differences in effective roughage level fed, and a variety of other factors. Overall, these results suggest that corn silage fed to growing calves should be kernel processed to enhance dry matter intake and daily weight gain, while kernel processed corn silage fed to finishing steers does not appreciably influence daily gain, efficiency of gain, or carcass parameters.

Risk-stratified faecal immunochemical testing (FIT) for urgent colonoscopy in Lynch syndrome during the COVID-19 pandemic.

BACKGROUND: Lynch syndrome is a hereditary cancer disease resulting in an increased risk of colorectal cancer. Herein, findings are reported from an emergency clinical service implemented during the COVID-19 pandemic utilizing faecal immunochemical testing ('FIT') in Lynch syndrome patients to prioritize colonoscopy while endoscopy services were limited. METHODS: An emergency service protocol was designed to improve colonoscopic surveillance access throughout the COVID-19 pandemic in England for people with Lynch syndrome when services were extremely restricted (1 March 2020 to 31 March 2021) and promoted by the English National Health Service. Requests for faecal immunochemical testing from participating centres were sent to the National Health Service Bowel Cancer Screening South of England Hub and a faecal immunochemical testing kit, faecal immunochemical testing instructions, paper-based survey, and pre-paid return envelope were sent to patients. Reports with faecal haemoglobin results were returned electronically for clinical action. Risk stratification for colonoscopy was as follows: faecal haemoglobin less than 10 µg of haemoglobin/g of faeces (µg/g)-scheduled within 6-12 weeks; and faecal haemoglobin greater than or equal to 10 µg/g-triaged via an urgent suspected cancer clinical pathway. Primary outcomes of interest included the identification of highest-risk Lynch syndrome patients and determining the impact of faecal immunochemical testing in risk-stratified colonoscopic surveillance. RESULTS: Fifteen centres participated from June 2020 to March 2021. Uptake was 68.8 per cent amongst 558 patients invited. For 339 eligible participants analysed, 279 (82.3 per cent) had faecal haemoglobin less than 10 µg/g and 60 (17.7 per cent) had faecal haemoglobin greater than or equal to 10 µg/g. In the latter group, the diagnostic accuracy of faecal immunochemical testing was 65.9 per cent and escalation to colonoscopy was facilitated (median 49 versus 122 days, χ2 = 0.0003, P < 0.001). CONCLUSION: Faecal immunochemical testing demonstrated clinical value for Lynch syndrome patients requiring colorectal cancer surveillance during the pandemic in this descriptive report of an emergency COVID-19 response service. Further longitudinal investigation on faecal immunochemical testing efficacy in Lynch syndrome is warranted and will be examined under the 'FIT for Lynch' study (ISRCTN15740250).

Proteomics dataset of epididymal fluid, seminal plasma, and proteins loosely attached to epididymal and ejaculated sperm from Angus bulls.

Peptides and proteins were identified by liquid chromatography with tandem mass spectrometry analysis (LCMS-MS) on an Orbitrap Velos mass spectrometer to further understand biological mechanisms that regulate increased longevity in epididymis compared to ejaculated sperm. Semen from sexually mature bulls were collected and then bulls were slaughtered to collect epididymal samples from the cauda epididymis. All samples were centrifuged to separate spermatozoa from fluids. A high ionic solution was used to remove surface proteins from spermatozoa. Four unique samples were generated: (1) epididymal fluid, (2) seminal plasma (ejaculated fluid), and proteins stripped from (3) epididymal sperm, and (4) ejaculated sperm. Samples were analyzed by LCMS-MS, and data were interpreted with Protein Pilot 5. False discovery rate (FDR) was set at 1%. Unique proteins (n = 458) were identified in ejaculated samples, 178 proteins in seminal plasma and 298 proteins stripped from ejaculated sperm. In epididymal samples, 311 proteins were identified in the fluid, and 334 were identified among proteins stripped from epididymal sperm. This dataset can be useful for further understand of biological mechanisms that control sperm longevity. This dataset is related to the article 'Proteomic analyses identify differences between bovine epididymal and ejaculated spermatozoa that contribute to longevity' by (Zoca et al., 2022).

Proteomic analyses identify differences between bovine epididymal and ejaculated spermatozoa that contribute to longevity.

Sperm are stored for extended periods of time in the epididymis, but upon ejaculation motility is increased and lifespan is decreased. The objective of this study was to identify differences in proteins between epididymis and ejaculated samples that are associated with longevity. Ejaculated semen was collected from mature Angus bulls (n = 9); bulls were slaughtered and epididymal semen was collected. Epididymal and ejaculated semen were centrifuged to separate sperm and fluid. Fluids were removed and sperm pellets were resuspended in a high ionic solution and vortexed to remove loosely attached proteins. Sperm samples were centrifuged, and the supernatant was removed; both fluid and sperm samples were snap frozen in liquid nitrogen and stored at -80 °C. Protein analysis was performed by LCMS/MS. A different group of yearling Angus cross bulls (n = 40) were used for sperm cultures. Ejaculated (n = 20) and epididymal (n = 20) semen were diluted and cultured in a commercial media at pH 5.8, 6.8 and 7.3, at 4 °C. Sperm were evaluated for motility and viability every 24 h until motility was lower than 20%. There was an effect of pH, time and pH by time interaction for motility and viability for both ejaculated and epididymal sperm (P ≤ 0.05). At 216 h of incubation epididymal sperm at pH 7.3 and ejaculated sperm at pH 6.8 reached motility below 20%. A total of 458 unique proteins were identified; 178, 298, 311, and 344 proteins were identified in ejaculated fluid, ejaculated sperm, epididymal fluid and epididymal sperm, respectively. There were 8, 24, 10, and 18 significant KEGG pathways (FDR <0.05) for ejaculated fluid, epididymal fluid, ejaculated sperm, and epididymal sperm, respectively. The metabolic pathway was identified as the most important KEGG pathway; glycolysis/gluconeogenesis, pentose phosphate, and glutathione metabolism pathways were significant among proteins only present in epididymal samples within the metabolic pathway. Other proteins identified that may be related to epididymal sperm's increased longevity were peroxidases and glutathione peroxidases for their antioxidant properties. In summary, energy metabolism in the epididymis appears to be more glycolytic compared to ejaculated and epididymis sperm have a larger number of antioxidants available which may be helping to maintain sperm in a quiescent state. Epididymal sperm remained viable (membrane integrity) longer than ejaculated sperm when cultured at the same pH.

Reduced trabecular bone mineral density and thinner cortices in men with distal forearm fractures.

Men with distal forearm fractures have reduced bone density, an increased risk of osteoporosis and of further fractures. The aim of the study was to investigate the structural determinants of these observations using quantitative CT (qCT). Ninety six men with low-trauma distal forearm fracture and 101 age-matched healthy control subjects were recruited. All subjects underwent a quantitative CT on a standard 64-slice whole body CT scanner. These were analysed on Mindways QCT PRO™ Software to generate volumetric and geometric data at the lumbar spine, femoral neck and total hip, ultra-distal and distal 33 % radius. Biochemical investigations, health questionnaires and measurements of bone turnover were made. Men with fracture had significantly lower total and trabecular vBMD at all sites. The greatest percentage reduction was at the ultra-distal radius (13.5 % total and 11.7 % trabecular vBMD). In the fracture group cortical vBMD was significantly higher in the femoral neck (p < 0.001) and maintained at the ultra-distal radius compared with control subjects. However, cortical cross-sectional area (CSA) and thickness were significantly reduced at the femoral neck (p < 0.001 and p = 0.002 respectively) and forearm sites (CSA ultradistal radius p = 0.001, cortical thickness p = 0.002, CSA distal one third radius p = 0.045 and cortical thickness p = 0.005). Cross sectional moment of inertia (CSMI) and section moduli were significantly reduced at the femoral neck (CSMI1 p = 0.002, CSMI2 p = 0.012 and section moduli Z1 p < 0.001, Z2 p = 0.004) and the ultra-distal radius (CSMI1 p = 0.008 and section moduli Z1 p = 0.018, Z2 p = 0.007). In stepwise logistic regression analysis distal forearm fracture showed the strongest association with a model comprising ultra-distal forearm trabecular vBMD (negative), procollagen type I N-terminal propeptide (PINP, positive) and sex hormone binding globulin (SHBG, negative). In conclusion, these observations explain the structural reasons for the increased fracture risk in men with distal forearm fractures.

Modulation of expression of estrus, steroidogenesis and embryo development following peri-artificial insemination nutrient restriction in beef heifers.

Nutritional changes immediately after insemination cause increased embryonic mortality, but the mechanisms controlling this are not well known. Our objective was to evaluate the impact of nutritional change on estrus expression, steroid concentrations, peripheral and uterine luminal fluid metabolites, and embryo quality in beef heifers. Heifers (n = 139) were assigned to one of two pre-artificial insemination (AI) dietary treatments: LOW (≤ 90% NEm) or HIGH (≥ 139% NEm). Heifers were on treatment for 33-36 days before AI (d0) when half of the heifers in each treatment were randomly reassigned to generate four treatments; HIGH-HIGH, HIGH-LOW, LOW-HIGH, and LOW-LOW. Heifers remained on treatments until embryo collection (d 6-8). Negative energy balance was achieved among LOW heifers as demonstrated by body weight loss and increased NEFA concentrations (P < 0.05). Pre-AI treatment influenced expression of estrus (P = 0.05; HIGH 80.4 ± 4.0% vs. LOW 69.4 ± 4.2%). Estradiol concentrations and interval to estrus were not affected by treatment (P > 0.55); however, progesterone concentrations were reduced among LOW compared to HIGH (3.57 ± 0.27, 4.64 ± 0.26 ng/mL, respectively; P = 0.004), and heifers maintained on the HIGH pre-AI diet had consistently greater concentrations of progesterone from d 0 to d 8 (P = 0.014). Pre-AI treatment influenced embryo stage (P = 0.05; HIGH 3.61 ± 0.32 vs. LOW 2.72 ± 0.30). Post-AI treatment affected embryo grade (P = 0.02; HIGH 1.78 ± 0.23 vs. LOW 2.64 ± 0.27). In summary, pre-AI nutrient restriction caused decreased expression of estrus, reduced progesterone concentrations after AI, and negatively impacted embryo development, while post-AI restriction hindered embryo quality.